Interacting proteins of protein kinase A regulatory subunit in Saccharomyces cerevisiae

F.GALELLO; MORENO, S; ROSSI, S

JOURNAL OF PROTEOMICS

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2014

1874-3919

ABSTRACT 28 29 cAMP-dependent protein kinase mediates many extracellular signals in eukaryotes. The 30 compartmentalization of PKA is an important level of control of the specificity of signal 31 transduction mediated by cAMP. Unlike mammalian PKA for which proof insights in 32 the mechanism that controls its localization through anchoring proteins (AKAPs) has 33 been obtained, in the case of Saccharomyces cerevisiae PKA there was little 34 information available. In this work, we present results that demonstrate the isolation and 35 identification of yeast PKA regulatory subunit (Bcy1) associated proteins using a MS36 based proteomic analysis and a bioinformatic approach. The verification of some of 37 these interactions was assessed by immunoprecipitation, pull down and co-localization 38 by subcellular fractionation. The key role of positively charged residues present in the 39 interaction domain of the identified proteins was demonstrated. The defined interaction 40 domain has therefore different molecular characteristics than conventional AKAP 41 domains. Finally we assess initial experiments to visualize the physiological relevance 42 of the interaction of both Ira2 and Hsp60 with Bcy1. Bcy1 interacts with Ira2 tethering 43 PKA to the Ras complex and Hsp60 chaperone localizes PKA to mitochondria and has a 44 role in the kinase stability.